Synergistic bacillus thuringiensis subsp. aizawai and chlorantraniliprole mixtures for diamondback moth, beet armyworm, soybean looper, corn earworm, cabbage looper, and southwestern corn borer control

ABSTRACT

The present invention generally relates to the use of synergistic amounts of  Bacillus thuringiensis  subsp.  aizawai  and chlorantraniliprole for the control of Diamondback Moth, Beet Armyworm, Soybean Looper, Corn Earworm, Cabbage Looper, and Southwestern Corn Borer. Specifically, the synergistic ratio of  Bacillus thuringiensis  subsp.  aizawai  to chlorantraniliprole is from about 1:0.00003 to about 1:3.

FIELD OF THE INVENTION

The present invention generally relates to the use of synergisticamounts of Bacillus thuringiensis subsp. aizawai and chlorantraniliprolefor the control of diamondback moth, beet armyworm, soybean looper, cornearworm, cabbage looper and southwestern corn borer.

BACKGROUND OF THE INVENTION

Lepidoptera is an order of insects which includes moths and butterflies.It is estimated that there are over 174,000 Lepidopteran species,included in an estimated 126 families. Lepidopteran species undergo acomplete metamorphosis during their life cycle. Adults mate and layeggs. The larvae that emerge from the eggs have a cylindrical body andchewing mouth parts. Larvae undergo several growth stages called instarsuntil they reach their terminal instar and then pupate. Lepidoptera thenemerge as adult butterflies or moths.

While some Lepidoptera species are generally considered beneficialorganisms due to their aesthetic appeal, many species cause devastatingdamage to crops. Specifically, diamondback moths, beet armyworms,soybean loopers, corn earworm, cabbage looper and southwestern cornborer are especially problematic to crop growers.

Diamondback moths (Plutella xylostella) are a widespread pest that candisperse long distances. Diamondback moth larvae eat the leaves, buds,flowers and seed-buds of cruciferous plants. A heavy infestation cancompletely remove all foliar tissue from a plant leaving only the leafveins. Even a lighter infestation can result in the unsuitability of anentire lot of produce for sale. In the past, diamondback moths have beentreated with a variety of insecticides including pyrethroids and otherinsecticides.

Beet armyworms (Spodoptera exigua) are another widespread pest that isdifficult to control. The larvae are voracious eaters that defoliatehost plants. Older instars can also burrow into the plants. The damageto the host plant renders it unmarketable. Beet armyworms are pests onnumerous types of crops.

Soybean loopers (Chrysodeixis includens) are a moth that is prevalent inNorth and South America. The larvae of soybean loopers can inflict heavyfoliage damage resulting in significant crop loss. Soybean loopers aredifficult to control with insecticides. Infestation of soybean looperscan be exacerbated after a non-selective insecticide removes the soybeanloopers' natural predators.

Corn earworms (Helicoverpa zea) have been referred to as the most costlycrop pest in the United States. Corn earworms are difficult to controlwith insecticides because they can burrow into the plants and avoidexposure to insecticide applications. Corn earworms have numerousnatural predators. However, predators and parasitoids alone are noteffective at preventing crop plant damage by Helicoverpa zea.

Cabbage loopers (Trichoplusia ni) are a destructive crop pest in NorthAmerica. During their larval stage, they eat three-times their bodyweight in plant material a day. Once they are established in a cropfield, they are difficult to control.

Southwestern corn borers (Diatraea grandiosella) cause crop damage inthe United States and Mexico. It is estimated that southwestern cornborers cause millions of dollars of damage each year. The first larvaethat emerge after overwintering feed on the whorl of the plant and cancause total destruction of the plant (dead heart). The second generationthat emerge later in the growing season feed on leaf axils andeventually bore into the stalks which can cause girdling and can alsolead to death of the plant.

Bacillus thuringiensis is a natural soil bacterium. Many Bacillusthuringiensis strains produce crystal proteins during sporulation calledδ-endotoxins which can be used as biological insecticides. Bacillusthuringiensis, subspecies aizawai, produces a crystal which paralyzesthe digestive system of some larvae within minutes. The larvaeeventually die off from the multiple deterious effects from toxininteractions with the gut tissues of the target pest. Bacillusthuringiensis subsp. aizawai is commercially available as XenTari®(available from Valent BioSciences Corporation, XenTari is a registeredtrademark of Valent BioSciences Corporation).

One advantage of using Bacillus thuringiensis subsp. aizawai is that itis target specific. It does not harm humans or other non-target species.Frequently when plants are treated with a non-selective insecticide, theinsecticide also kills natural predators of other pests. This can causea rebound effect in the target insect or other opportunistic pestspecies. For example, after applying a non-selective pesticide to killcorn borers, a spider mite infestation might occur because thenon-selective pesticide also killed the spider mites' natural predators.

Yet another advantage of Bacillus thuringiensis subsp. aizawai is thatit can be used on organic crops. With no mandated pre-harvest interval,it can also be used on crops right before harvest. This provides organicgrowers, who have few options for pest control, a safe and effective wayto manage insect infestations that could ultimately ruin an entire crop.

Chlorantraniliprole is an anthranilic diamide. Chlorantraniliprole haslow toxicity to humans and mammals. Further, it is effective at low userates. Like Bacillus thuringiensis subsp. aizawai, chlorantraniliprolemust be eaten by larvae in order to be effective. Chlorantraniliproleforces muscles within the larvae to release all of their stored calcium,causing the larvae to stop eating and eventually die.Chlorantraniliprole is commercially available, for example, as Coragen®(available from Dupont™, Coragen is a registered trademark of E. I. duPont de Nemours and Company).

Wakil, et al., applied Bacillus thuringiensis kurstaki andchlorantraniliprole to Helicoverpa armigera species (Wakil, et al.,Effects of Interactions Among Metarhizium anisopliae, BacillusThuringiensis and Chlorantraniliprole on the Mortality and Pupation ofSix Geographically Distinct Helicoverpa armigera Field Populations,Phytoparasitica, 2013, 21:221-234). Helicoverpa is another one of the126 families of Lepidoptera. Based on the results on Helicoverpaarmigera, one of skill in the art would not have been able to predicthow any of the 174,000+ Lepidoptera species would respond to thetreatment with a different subspecies of Bacillus thuringiensis,Bacillus thuringiensis subsp. aizawai, with chlorantraniliprole.

Accordingly, there is a need for safe and effective ways to controldiamondback moth, beet armyworm, soybean looper, corn earworm, cabbagelooper and southwestern corn borer. These methods should be easy toapply, have increased efficacy, and be cost effective.

SUMMARY OF THE INVENTION

The present invention is directed to methods for controlling diamondbackmoth (Plutella xylostella), beet armyworm (Spodoptera exigua), soybeanlooper (Chrysodeixis includens), corn earworm (Helicoverpa zea), cabbagelooper (Trichoplusia ni), and southwestern corn borer (Diatraeagrandiosella) comprising applying a synergistic amount of Bacillusthuringiensis subsp. aizawai and chlorantraniliprole to a plant, whereinthe ratio of Bacillus thuringiensis subsp. aizawai tochlorantraniliprole is from about 1:0.00003 to about 1:3.

DETAILED DESCRIPTION OF THE INVENTION

Applicant discovered that the use of Bacillus thuringiensis subsp.aizawai and chlorantraniliprole at a ratio range of from about 1:0.00003to about 1:3 provided unexpected synergistic effects against specificLepidopteran species (compare Examples 1 to 3 and 5 to 7 with Example 4below). This synergy was unexpected because the species' responses tosimilar treatments has been unpredictable and the species' responses tothe treatments herein were varied. Although the use of Bacillusthuringiensis subsp. aizawai and chlorantraniliprole was not synergisticfor sugarcane borer, Applicant was surprised to discover that thesynergistic ratios of the present invention provided control ofdiamondback moth, beet armyworm, soybean looper, corn earworm, cabbagelooper and southwestern corn borer.

The Bacillus thuringiensis subsp. aizawai and chlorantraniliprolesynergistic mixtures are also safe to use on edible plants. Further, thecomponents of the mixtures are target specific and pose low to no riskto beneficial insects or animals.

Another advantage of the present invention is that the combination ofBacillus thuringiensis subsp. aizawai and chlorantraniliprole alignswith Integrated Pest Management (IPM) principles and will reduce theability of the insects to develop resistance to chlorantraniliprole. Bycombining two different products with different modes of action, theability of the insects to dominantly express mutations which overcomeboth Bacillus thuringiensis subsp. aizawai toxins andchlorantraniliprole is very unlikely. This means that the mixture ofBacillus thuringiensis subsp. aizawai and chlorantraniliprole can beapplied repeatedly in the same season and year after year with minimalrisk of resistance developing.

Yet another advantage of the present invention is that it allows forless chlorantraniliprole and less Bacillus thuringiensis subsp. aizawaito be applied to the plant. For example, within label rates, sub-lethaldoses of each can be applied to achieve a lethal dose and control of thelarvae. This allows for a significant cost saving to the grower.

A further advantage is that Bacillus thuringiensis subsp. aizawai andchlorantraniliprole are target-specific. This means that humans andother, non-target organisms—such as natural predators of diamondbackmoth, beet armyworm, soybean looper, corn earworm, cabbage looper, andsouthwestern corn borer—will not be harmed by the methods of the presentinvention.

In an embodiment, the present invention is directed to methods forcontrolling a crop plant pest selected from the group consisting ofdiamondback moth (Plutella xylostella), beet armyworm (Spodopteraexigua), soybean looper (Chrysodeixis includens), corn earworm(Helicoverpa zea), cabbage looper (Trichoplusia ni), and southwesterncorn borer (Diatraea grandiosella) comprising applying a synergisticamount of Bacillus thuringiensis subsp. aizawai and chlorantraniliproleto a plant, wherein the ratio of Bacillus thuringiensis subsp. aizawaito chlorantraniliprole is from about 1:0.00003 to about 1:3.

As used herein, “crop plant pest” only refers to diamondback moth(Plutella xylostella), beet armyworm (Spodoptera exigua), soybean looper(Chrysodeixis includens), corn earworm (Helicoverpa zea), cabbage looper(Trichoplusia ni), and southwestern corn borer (Diatraea grandiosella)

In a preferred embodiment, the ratio of Bacillus thuringiensis subsp.aizawai to chlorantraniliprole is from about 1:0.001 to about 1:1. In amore preferred embodiment, the ratio of Bacillus thuringiensis subsp.aizawai to chlorantraniliprole is from about 1:0.04 to about 1:0.8.

In another embodiment, the present invention is directed to methods forcontrolling a crop plant pest wherein the amount of Bacillusthuringiensis subsp. aizawai is from about 50 to about 4,000 grams perhectare. In a preferred embodiment, the amount of Bacillus thuringiensissubsp. aizawai is from about 100 to about 1,300 grams per hectare. In amore preferred embodiment, the amount of Bacillus thuringiensis subsp.aizawai is from about 150 to about 1,250 grams per hectare.

In a further embodiment, the present invention is directed to methodsfor controlling a crop plant pest wherein the amount of Bacillusthuringiensis subsp. aizawai is from about 7,000 to about 200,000 IU/mg.In a preferred embodiment, the amount of Bacillus thuringiensis subsp.aizawai is from about 20,000 to about 170,000 IU/mg. In a more preferredembodiment, the amount of Bacillus thuringiensis subsp. aizawai is fromabout 25,000 to about 100,000 IU/mg.

In yet another embodiment, the present invention is directed to methodsfor controlling a crop plant pest wherein the amount of Bacillusthuringiensis subsp. aizawai is from about 5,000 to about 100,000Spodoptera U/mg. In a preferred embodiment, the amount of Bacillusthuringiensis subsp. aizawai is from about 20,000 to about 90,000Spodoptera U/mg. In a more preferred embodiment, the amount of Bacillusthuringiensis subsp. aizawai is from about 40,000 to about 70,000Spodoptera U/mg.

Although in some embodiments, the rates of Bacillus thuringiensis subsp.aizawai are expressed in grams/hectare, IU/mg, or Spodoptera U/mg, theinvention is not limited to these methods of measuring potency. If otherproducts are developed or marketed with other potency measurements, itis within the knowledge of one of skill in the art, based on Applicant'steaching herein, to convert the rates to effective amounts consistentwith the invention herein to achieve synergistic control of the targetcrop plant pest.

Further, the present invention is not limited to a specific type offormulation. For example, in the examples herein, a dry flowablegranular formulation was used as the source of Bacillus thuringiensissubsp. aizawai. However, other types of formulations may be used,including but not limited to wettable powder formulations, waterdispersible granules, granules, and emulsifiable suspensionconcentrates. Technical grade powders may also be used.

Suitable Bacillus thuringiensis subsp. aizawai subspecies strainsinclude, but are not limited to, VBTS-1857, GB413, GC-91, andtransconjugated, recombinant and/or genetically engineered subspeciesthereof.

Suitable Bacillus thuringiensis subsp. aizawai commercial productsinclude, but are not limited to, XenTari® (as indicated above, availablefrom Valent BioSciences Corporation, XenTari is a registered trademarkof Valent BioSciences Corporation), Solbit (available from Green BiotechCompany), Bacchus® (available from Certis, Bacchus is registeredtrademark of Certis USA, L.L.C.), Agree (available from Certis, Agree isregistered trademark of Certis USA, L.L.C.), Jackpot® (available fromCertis, Jackpot is registered trademark of Certis USA, L.L.C.), andTurex® (available from Certis, Turex is registered trademark of CertisUSA, L.L.C.).

In yet another embodiment, the present invention is directed to methodsfor controlling a crop plant pest wherein the amount ofchlorantraniliprole is from about 20 to about 150 grams per hectare. Ina preferred embodiment, the amount of chlorantraniliprole is from about30 to about 130 grams per hectare. In a more preferred embodiment, theamount of chlorantraniliprole is from about 50 to about 110 grams perhectare.

The examples herein used a commercial product of chlorantraniliprole butthe invention is not limited to the use of this commercial product.Suitable chlorantraniliprole products include, but are not limited to,Coragen® (as indicated above, available from Dupont™, Coragen is aregistered trademark of E. I. du Pont de Nemours and Company),Acelepryn™ (available from Dupont™), and Rynaxypyr® (also available fromDupont™, Rynaxypyr is a registered trademark of E. I. du Pont de Nemoursand Company).

In a further embodiment, the present invention is directed to methodsfor controlling a crop plant pest comprising applying a synergisticamount of Bacillus thuringiensis subsp. aizawai and chlorantraniliproleto a plant, wherein the ratio of Bacillus thuringiensis subsp. aizawaito chlorantraniliprole is from about 1:0.00003 to about 1:3, and whereinthe plant is selected from the group consisting of root and tubervegetables, bulb vegetables, leafy non-brassica vegetables, leafybrassica vegetables, succulent or dried legumes, fruiting vegetables,cucurbit vegetables, citrus fruits, pome fruits, stone fruits, berry andsmall fruits, tree nuts, cereal grains, forage and fodder grasses andhay, non-grass animal feeds, herbs, spices, flowers, bedding plants,ornamental flowers, artichoke, asparagus, coffee, cotton, tropicalfruits, hops, malanga, peanut, pomegranate, oil seed vegetables,sugarcane, tobacco, turf, and watercress.

In another embodiment, the crop plant is genetically modified. A“genetically modified” crop plant is one that has had specific genesremoved, modified or additional gene copies of native or foreign DNA.The change in the crop plant's DNA may result in can result in changesin the type or amount of RNA, proteins and/or other molecules that thecrop plant produces which may affect its response to abiotic (e.g.herbicide) or biotic (e.g. insects) stresses, and/or affect its growth,development, or yield.

When Bacillus thuringiensis subsp. aizawai and chlorantraniliprole areapplied to cotton to control beet armyworm, the most preferred rate ofBacillus thuringiensis subsp. aizawai is from about 600 to about 1,250grams per hectare. When Bacillus thuringiensis subsp. aizawai andchlorantraniliprole are applied to cotton to control beet armyworm, themost preferred rate of chlorantraniliprole is from about 50 to about 110grams per hectare. Accordingly, when Bacillus thuringiensis subsp.aizawai and chlorantraniliprole are applied to cotton to control beetarmyworm, the most preferred ratio of Bacillus thuringiensis subsp.aizawai to chlorantraniliprole is from about 1:0.04 to about 1:0.2.

When Bacillus thuringiensis subsp. aizawai and chlorantraniliprole areapplied to any crop except cotton to control beet armyworm, the mostpreferred rate of Bacillus thuringiensis subsp. aizawai is from about600 to about 1,250 grams per hectare. When Bacillus thuringiensis subsp.aizawai and chlorantraniliprole are applied to any crop except cotton tocontrol beet armyworm, the most preferred rate of chlorantraniliprole isfrom about 50 to about 75 grams per hectare. Accordingly, when Bacillusthuringiensis subsp. aizawai and chlorantraniliprole are applied to anycrop except cotton to control beet armyworm, the most preferred ratio ofBacillus thuringiensis subsp. aizawai to chlorantraniliprole is fromabout 1:0.04 to about 1:0.15.

When Bacillus thuringiensis subsp. aizawai and chlorantraniliprole areapplied to any crop to control diamondback moth, corn earworm, cabbagelooper, and/or southwestern corn borer the most preferred rate ofBacillus thuringiensis subsp. aizawai is from about 150 to about 1,250grams per hectare. When Bacillus thuringiensis subsp. aizawai andchlorantraniliprole are applied to any crop to control diamondback moth,and/or corn earworm, the most preferred rate of chlorantraniliprole isfrom about 50 to about 75 grams per hectare. Accordingly, when Bacillusthuringiensis subsp. aizawai and chlorantraniliprole are applied to anycrop to control diamondback moth, and/or corn earworm, the mostpreferred ratio of Bacillus thuringiensis subsp. aizawai tochlorantraniliprole is from about 1:0.04 to about 1:0.05.

When Bacillus thuringiensis subsp. aizawai and chlorantraniliprole areapplied to cotton to control soybean looper, the most preferred rate ofBacillus thuringiensis subsp. aizawai is from about 150 to about 1,250grams per hectare. When Bacillus thuringiensis subsp. aizawai andchlorantraniliprole are applied to cotton to control soybean looper, themost preferred rate of chlorantraniliprole is from about 50 to about 110grams per hectare. Accordingly, when Bacillus thuringiensis subsp.aizawai and chlorantraniliprole are applied to cotton to control soybeanlooper, the most preferred ratio of Bacillus thuringiensis subsp.aizawai to chlorantraniliprole is from about 1:0.04 to about 1:0.8.

When Bacillus thuringiensis subsp. aizawai and chlorantraniliprole areapplied to any crop except cotton to control soybean looper, the mostpreferred rate of Bacillus thuringiensis subsp. aizawai is from about150 to about 1,250 grams per hectare. When Bacillus thuringiensis subsp.aizawai and chlorantraniliprole are applied to any crop except cotton tocontrol soybean looper, the most preferred rate of chlorantraniliproleis from about 50 to about 75 grams per hectare. Accordingly, whenBacillus thuringiensis subsp. aizawai and chlorantraniliprole areapplied to any crop except cotton to control soybean looper, the mostpreferred ratio of Bacillus thuringiensis subsp. aizawai tochlorantraniliprole is from about 1:0.04 to about 1:0.5.

In a preferred embodiment, the root and tuber vegetables are selectedfrom the group consisting of arracacha, arrowroot, Chinese artichoke,Jerusalem artichoke, garden beet, sugar beet, edible burdock, ediblecanna, carrot, bitter cassava, sweet cassava, celeriac, root chayote,turnip-rooted chervil, chicory, chufa, dasheen (taro), ginger, ginseng,horseradish, leren, turnip-rooted parsley, parsnip, potato, radish,oriental radish, rutabaga, salsify, black salsify, Spanish salsify,skirret, sweet potato, tanier, turmeric, turnip, yam bean, true yam, andcultivars, varieties and hybrids thereof.

In another preferred embodiment, the bulb vegetables are selected fromthe group consisting of fresh chive leaves, fresh Chinese chive leaves,bulb daylily, elegans hosta, bulb fritillaria, fritillaria leaves, bulbgarlic, great-headed bulb garlic, serpent bulb garlic, kurrat, lady'sleek, leek, wild leek, bulb lily, Beltsville bunching onion, bulb onion,Chinese bulb onion, fresh onion, green onion, macrostem onion, pearlonion, potato bulb onion, potato bulb, tree onion tops, Welsh oniontops, bulb shallot, fresh shallot leaves, and cultivars, varieties andhybrids thereof.

In a further embodiment, the leafy non-brassica vegetables are selectedfrom the group consisting of Chinese spinach Amaranth, leafy Amaranth,arugula (roquette), cardoon, celery, Chinese celery, celtuce, chervil,edible-leaved chrysanthemum, garland chrysanthemum, corn salad, gardencress, upland cress, dandelion, dandelion leaves, sorrels (dock), endive(escarole), Florence fennel, head lettuce, leaf lettuce, orach, parsley,garden purslane, winter purslane, radicchio (red chicory), rhubarb,spinach, New Zealand spinach, vine spinach, Swiss chard, Tampala, andcultivars, varieties and hybrids thereof.

In another embodiment, the leafy brassica vegetables are selected fromthe group consisting of broccoli, Chinese broccoli (gai lon), broccoliraab (rapini), Brussels sprouts, cabbage, Chinese cabbage (bok choy),Chinese napa cabbage, Chinese mustard cabbage (gai choy), cauliflower,cavalo broccoli, collards, kale, kohlrabi, mizuna, mustard greens,mustard spinach, rape greens, and cultivars, varieties and hybridsthereof.

In yet another embodiment, the succulent or dried vegetable legumes areselected from the group consisting of Lupinus beans, Phaseolus beans,Vigna beans, broad beans (fava), chickpea (garbanzo), guar, jackbean,lablab bean, lentil, Pisum peas, pigeon pea, soybean, immature seedsoybean, sword bean, peanut, and cultivars, varieties and hybridsthereof. In a preferred embodiment, the Lupinus beans include grainlupin, sweet lupin, white lupin, white sweet lupin, and hybrids thereof.In another preferred embodiment, the Phaseolus beans include field bean,kidney bean, lima bean, navy bean, pinto bean, runner bean, snap bean,tepary bean, wax bean, and hybrids thereof. In yet another preferredembodiment, the Vigna beans include adzuki bean, asparagus bean,blackeyed bean, catjang, Chinese longbean, cowpea, Crowder pea, mothbean, mung bean, rice bean, southern pea, urd bean, yardlong bean, andhybrids thereof. In another embodiment, the Pisum peas include dwarfpea, edible-podded pea, English pea, field pea, garden pea, green pea,snow pea, sugar snap pea, and hybrids thereof. In a preferredembodiment, the dried vegetable legume is soybean. In a more preferredembodiment, the dried vegetable legume is genetically modified soybean.

In a further embodiment, the fruiting vegetables are selected from thegroup consisting of bush tomato, cocona, currant tomato, gardenhuckleberry, goji berry, groundcherry, martynia, naranjilla, okra, peaeggplant, pepino, peppers, non-bell peppers, Scout tomato fieldsroselle, eggplant, scarlet eggplant, African eggplant, sunberry,tomatillo, tomato, tree tomato, and cultivars, varieties and hybridsthereof. In a preferred embodiment, the peppers include bell peppers,chili pepper, cooking pepper, pimento, sweet peppers, and hybridsthereof.

In an embodiment, the cucurbit vegetables are selected from the groupconsisting of Chayote, Chayote fruit, waxgourd (Chinese preservingmelon), citron melon, cucumber, gherkin, edible gourds, Momordicaspecies, muskmelons, pumpkins, summer squashes, winter squashes,watermelon, and cultivars, varieties and hybrids thereof. In a preferredembodiment, edible gourds include hyotan, cucuzza, hechima, Chineseokra, and hybrids thereof. In another preferred embodiment, theMomordica vegetables include balsam apple, balsam pear, bittermelon,Chinese cucumber, and hybrids thereof. In another preferred embodiment,the muskmelon include true cantaloupe, cantaloupe, casaba, crenshawmelon, golden pershaw melon, honeydew melon, honey balls, mango melon,Persian melon, pineapple melon, Santa Claus melon, snake melon, andhybrids thereof. In yet another preferred embodiment, the summer squashinclude crookneck squash, scallop squash, straightneck squash, vegetablemarrow, zucchini, and hybrids thereof. In a further preferredembodiment, the winter squash includes butternut squash, calabaza,hubbard squash, acorn squash, spaghetti squash, and hybrids thereof.

In another embodiment, the citrus fruits are selected from the groupconsisting of limes, calamondin, citron, grapefruit, Japanese summergrapefruit, kumquat, lemons, Mediterranean mandarin, sour orange, sweetorange, pummel, Satsuma mandarin, tachibana orange, tangelo, mandarintangerine, tangor, trifoliate orange, uniq fruit, and cultivars,varieties and hybrids thereof. In a preferred embodiment, the limes areselected from the group consisting of Australian desert lime, Australianfinger lime, Australian round lime, Brown River finger lime, mount whitelime, New Guinea wild lime, sweet lime, Russell River lime, Tahiti lime,and hybrids thereof.

In an embodiment, the pome fruits are selected from the group consistingof apple, azarole, crabapple, loquat, mayhaw, medlar, pear, Asian pear,quince, Chinese quince, Japanese quince, tejocote, and cultivars,varieties and hybrids thereof

In another embodiment, the stone fruits are selected from the groupconsisting of apricot, sweet cherry, tart cherry, nectarine, peach,plum, Chicksaw plum, Damson plum, Japanese plum, plumcot, fresh prune,and cultivars, varieties and hybrids thereof.

In a further embodiment, the berries and small fruits are selected fromthe group consisting of Amur river grape, aronia berry, bayberry,bearberry, bilberry, blackberry, blueberry, lowbush blueberry, highbushblueberry, buffalo currant, buffaloberry, che, Chilean guava,chokecherry, chokeberry, cloudberry, cranberry, highbush cranberry,black currant, red currant, elderberry, European barberry, gooseberry,grape, edible honeysuckle, huckleberry, jostaberry, Juneberry (Saskatoonberry), lingonberry, maypop, mountain pepper berries, mulberry,muntries, native currant, partridgeberry, phalsa, pincherry, blackraspberry, red raspberry, riberry, salal, schisandra berry, seabuckthorn, serviceberry, strawberry, wild raspberry, and cultivars,varieties and hybrids thereof. In a preferred embodiment, theblackberries include Andean blackberry, arctic blackberry, bingleberry,black satin berry, boysenberry, brombeere, California blackberry,Chesterberry, Cherokee blackberry, Cheyenne blackberry, commonblackberry, coryberry, darrowberry, dewberry, Dirksen thornless berry,evergreen blackberry, Himalayaberry, hullberry, lavacaberry, loganberry,lowberry, Lucreliaberry, mammoth blackberry, marionberry, mora, muresderonce, nectarberry, Northern dewberry, olallieberry, Oregon evergreenberry, phenomenalberry, rangeberry, ravenberry, rossberry, Shawneeblackberry, Southern dewberry, tayberry, youngberry, zarzamora, andhybrids thereof.

In another embodiment, the tree nuts are selected from the groupconsisting of almond, beech nut, Brazil nut, butternut, cashew,chestnut, chinquapin, hazelnut (filbert), hickory nut, macadamia nut,pecan, pistachio, black walnut, English walnut, and cultivars, varietiesand hybrids thereof.

In a further embodiment, the cereal grains are selected from the groupconsisting of barley, buckwheat, pearl millet, proso millet, oats, corn,field corn, sweet corn, seed corn, popcorn, rice, rye, sorghum (milo),sorghum species, grain sorghum, sudangrass (seed), teosinte, triticale,wheat, wild rice, and cultivars, varieties and hybrids thereof. In apreferred embodiment, the cereal grain is corn. In a more preferredembodiment, the cereal grain is genetically modified corn.

In yet another embodiment, the grass forage, fodder and hay are selectedfrom the group consisting of grasses that are members of the Gramineaefamily except sugarcane and those species included in the cereal grainsgroup, pasture and range grasses, and grasses grown for hay or silage.In further embodiments, the Gramineae grasses may be green or cured.

In an embodiment, the non-grass animal feeds are selected from the groupconsisting of alfalfa, velvet bean, trifolium clover, melilotus clover,kudzu, lespedeza, lupin, sainfoin, trefoil, vetch, crown vetch, milkvetch, and cultivars, varieties and hybrids thereof.

In another embodiment, the herbs and spices are selected from the groupconsisting of allspice, angelica, anise, anise seed, star anise, annattoseed, balm, basil, borage, burnet, chamomile, caper buds, caraway, blackcaraway, cardamom, cassia bark, cassia buds, catnip, celery seed,chervil, chive, Chinese chive, cinnamon, clary, clove buds, corianderleaf, coriander seed, costmary, culantro leaves, culantro seed, cilantroleaves, cilantro seed, cumin, dillweed, dill seed, fennel, commonfennel, Florence fennel seed, fenugreek, grains of paradise, horehound,hyssop, juniper berry, lavender, lemongrass, leaf lovage, seed lovage,mace, marigold, marjoram, mint, mustard seed, nasturtium, nutmeg,parsley, pennyroyal, black pepper, white pepper, poppy seed, rosemary,rue, saffron, sage, summer savory, winter savory, sweet bay, tansy,tarragon, thyme, vanilla, wintergreen, woodruff, wormwood, andcultivars, varieties and hybrids thereof. In a preferred embodiment, themints are selected from the group consisting of spearmint, peppermint,and hybrids thereof.

In yet another embodiment, artichokes are selected from the groupconsisting of Chinese artichoke, Jerusalem artichoke, and cultivars,varieties and hybrids thereof.

In an embodiment, the tropical fruits are selected from the groupconsisting of anonna, avocado, fuzzy kiwifruit, hardy kiwifruit, banana,plantain, caimito, carambola (star fruit), guava, longan, sapodilla,papaya, passion fruit, mango, lychee, jackfruit, dragon fruit, mameysapote, coconut cherimoya, canistrel, monster, wax jambu, pomegranate,rambutan, pulasan, Pakistani mulberry, langsat, chempedak, durian, figpineapple, jaboticaba, mountain apples, pineapple, and cultivars,varieties and hybrids thereof.

In a further embodiment, the oil seed vegetables are selected from thegroup consisting of borage, calendula, castor oil plant, tallowtree,cottonseed, crambe, cuphea, echium, euphorbia, evening primrose, flaxseed, gold of pleasure, hare's ear, mustard, jojoba, lesquerella,lunaria, meadowfoam, milkweed, niger seed, oil radish, poppy seed,rosehip, sesame, stokes aster, sweet rocket, tallowwood, tea oil plant,vermonia, canola, or oil rapeseed, safflower, sunflower, and cultivars,varieties and hybrids thereof.

The synergistic amounts of Bacillus thuringiensis subsp. aizawai andchlorantraniliprole may be applied to seeds, foliage, or an area where aplant is intended to grow.

The synergistic amounts of Bacillus thuringiensis subsp. aizawai andchlorantraniliprole may be applied once or many times during a growingseason. If Bacillus thuringiensis subsp. aizawai and chlorantraniliproleare applied more than one time, the total amount applied should notexceed a yearly maximum rate as determined by environmental protectionagencies.

As used herein, “plant” refers to at least one plant and not a plantpopulation.

As used herein, “control” or “controlling” means a decline in the amountof damage to the plants from the larvae, reduction of pest population,interference with life cycle development or other physiological orbehavioral effect that results in plant protection.

As used herein, all numerical values relating to amounts, weightpercentages and the like, are defined as “about” or “approximately” eachparticular value, plus or minus 10%. For example, the phrase “at least5.0% by weight” is to be understood as “at least 4.5% to 5.5% byweight.” Therefore, amounts within 10% of the claimed values areencompassed by the scope of the claims.

The disclosed embodiments are simply exemplary embodiments of theinventive concepts disclosed herein and should not be considered aslimiting, unless so stated.

The following examples are intended to illustrate the present inventionand to teach one of ordinary skill in the art how to make and use theinvention. They are not intended to be limiting in any way.

EXAMPLES

The following examples illustrate the synergy of Bacillus thuringiensissubsp. aizawai and chlorantraniliprole when controlling diamondbackmoth, beet armyworm, soybean looper, corn earworm, cabbage looper, andsouthwestern corn borer. Xentari® was used as the source of Bacillusthuringiensis subsp. aizawai and Coragen® was used as the source ofchlorantraniliprole. The present invention is not limited to theproducts or formulation types used herein. In each example below, thestudies were conducted as follows.

For these tests, standardized laboratory leaf dip methods were used toinoculate plant material with treatment(s). Dry, treated leaves wereplaced into Petri dishes (100×25 mm) containing filter paper wetted with500 μl of distilled H₂O (“dH₂O”). Each dish was then infested withbetween 5 and 10 larvae, dependent on species. Efficacy ratings weretaken at specified intervals. Synergy ratings were calculated for eachtest.

Example 1 Diamondback Moth

In this study, the response of diamondback moth larvae to synergisticamounts of Bacillus thuringiensis subsp. aizawai (“Bta”) andchlorantraniliprole was observed. The results of this study can be seenbelow in Table 1.

TABLE 1 Time after % Efficacy treat- Neg. Bta + ment Controlchlorantraniliprole Synergy (h) dH₂O Bta Chlorantraniliprole (Ratio1:0.0003) Ratio 24 0 7 7 13 0.96 48 0 10 7 43 2.64

As seen in Table 1, the mixtures of the present invention provided amore than additive effect. By using the following formula, Applicant wasable to determine that this response was synergistic: %Cexp=A+B−(AB/100).

% Cexp=A+B−(AB/100), where % Cexp is the expected efficacy and “in whichA and B are the control levels given by the single [insecticides]. Ifthe ratio between the experimentally observed efficacy of the mixtureC_(obs) and the expected efficacy of the mixture is greater than 1,synergistic interactions are present in the mixture.” (Gisi,Synergisitic Interaction of Fungicides in Mixtures, The AmericanPhytopathological Society, 86:11, 1273-1279, 1996). Adopting aconservative approach, Applicant determined synergy to be present atratios of ≧1.15.

Bacillus thuringiensis subsp. aizawai was applied at a concentration of2.7 ppm (2.7 μg/ml). Chlorantraniliprole was applied at a concentrationof 0.0009 ppm (0.0009 μg/ml). The Bacillus thuringiensis subsp.aizawai/chlorantraniliprole mixture was applied at a concentration of2.7 ppm Bacillus thuringiensis subsp. aizawai and 0.0009 ppmchlorantraniliprole.

In order to determine synergy, rates below normal field rate ranges mustbe used. If normal field rate ranges are used, all of the larvae woulddie (combining a lethal or near lethal dose of Bacillus thuringiensissubsp. aizawai with a lethal dose of chlorantraniliprole would mostlikely lead to larvae death) in every treatment and synergy would not beable to be determined. A synergy ratio that is indicative of synergy isthis assay is a predictor of the synergy that will be seen in the fieldat normal field rates (or at rates that occur naturally as the activeingredients are degraded over time by exposure to rain, UV radiation,and temperature extremes). This assay was chosen for its ability toaccurately predict mortality rates of larvae in the field.

The results of this calculation indicated that the synergy ratio was0.96 at 24 hours and 2.64 at 48 hours. As a finding of higher than 1 isindicative of synergy (per Gisi, or even≧1.15 per Applicant), the ratioof 2.64 is synergistic. Synergy was shown at a ratio of Bacillusthuringiensis subsp. aizawai to chlorantraniliprole of 1:0.0003.

Example 2 Beet Armyworm

In this study, the response of beet armyworm larvae to amounts ofBacillus thuringiensis subsp. aizawai and chlorantraniliprole wasobserved. The results of this study can be seen in Table 2.

TABLE 2 Time after % Efficacy treat- Neg. Bta + ment Controlchlorantraniliprole Synergy (h) dH₂O Bta Chlorantraniliprole (Ratio1:0.0003) Ratio 24 0 3 10 17 1.34 48 0 10 13 27 1.24

Bacillus thuringiensis subsp. aizawai was applied at a concentration of2.7 ppm (2.7 μg/ml). Chlorantraniliprole was applied at a concentrationof 0.0009 ppm (0.0009 μg/ml). The Bacillus thuringiensis subsp.aizawai/chlorantraniliprole mixture was applied at a concentration of2.7 ppm Bacillus thuringiensis subsp. aizawai and 0.0009 ppmchlorantraniliprole.

As seen in Table 2, the mixtures of the present invention provided amore than additive effect. By using the following formula, Applicant wasable to determine that this response was synergistic: %Cexp=A+B−(AB/100).

The results of this calculation indicated that the synergy ratio was1.34 at 24 hours and 1.24 at 48 hours. As a finding of higher than 1 isindicative of synergy, the ratio of 1.24 is synergistic. Synergy wasshown at a ratio of Bacillus thuringiensis subsp. aizawai tochlorantraniliprole of 1:0.0003.

Example 3 Cabbage Looper

In this study, the response of cabbage looper larvae to amounts ofBacillus thuringiensis subsp. aizawai and chlorantraniliprole wasobserved. The results of this study are below in Table 3.

TABLE 3 Time after % Efficacy treat- Neg. Bta + ment Controlchlorantraniliprole Synergy (h) dH₂O Bta Chlorantraniliprole (Ratio1:0.0003) Ratio 24 0 7 7 13 0.96 48 0 7 10 23 1.41

Bacillus thuringiensis subsp. aizawai was applied at a concentration of2.7 ppm (2.7 μg/ml). Chlorantraniliprole was applied at a concentrationof 0.0009 ppm (0.0009 μg/ml). The Bacillus thuringiensis subsp.aizawai/chlorantraniliprole mixture was applied at a concentration of2.7 ppm Bacillus thuringiensis subsp. aizawai and 0.0009 ppmchlorantraniliprole.

As seen in Table 3, the mixtures of the present invention provided amore than additive effect. By using the following formula, Applicant wasable to determine that this response was synergistic: %Cexp=A+B−(AB/100).

The results of this calculation indicated that the synergy ratio was0.96 at 24 hours and 1.41 at 48 hours. As a finding of higher than 1 isindicative of synergy, the ratio of 1.41 is synergistic. Synergy wasshown at a ratio of Bacillus thuringiensis subsp. aizawai tochlorantraniliprole of 1:0.0003.

Example 4 Sugarcane Borer

In this study, the response of sugarcane borer larvae to amounts ofBacillus thuringiensis subsp. aizawai and chlorantraniliprole wasobserved. The results of this study are in Table 4.

TABLE 4 Time after % Efficacy treat- Neg. Bta + ment Controlchlorantraniliprole Synergy (h) dH₂O Bta Chlorantraniliprole (Ratio1:0.0003) Ratio 24 0 3 6 3 No Synergy 48 0 9 12 15 No Synergy

Bacillus thuringiensis subsp. aizawai was applied at a concentration of27 ppm (27 μg/ml). Chlorantraniliprole was applied at a concentration of0.0009 ppm (0.0009 μg/ml). The Bacillus thuringiensis subsp.aizawai/chlorantraniliprole mixture was applied at a concentration of 27ppm Bacillus thuringiensis subsp. aizawai and 0.0009 ppmchlorantraniliprole.

As seen in Table 4, the mixtures of the present invention failed toprovide more than an additive effect. By using the following formula,Applicant was able to determine that this response was not synergistic:% Cexp=A+B−(AB/100).

Example 5 Southwestern Corn Borer

In this study, the response of southwestern corn borer larvae to amountsof Bacillus thuringiensis subsp. aizawai and chlorantraniliprole wasobserved. The results of this study are in Table 5.

TABLE 5 Time after % Efficacy treat- Neg. Bta + ment Controlchlorantraniliprole Synergy (h) dH₂O Bta Chlorantraniliprole (Ratio1:0.0003) Ratio 24 0 6 3 19 2.15 48 0 10 7 19 1.17

Bacillus thuringiensis subsp. aizawai was applied at a concentration of2.7 ppm (2.7 μg/ml). Chlorantraniliprole was applied at a concentrationof 0.0009 ppm (0.0009 μg/ml). The Bacillus thuringiensis subsp.aizawai/chlorantraniliprole mixture was applied at a concentration of2.7 ppm Bacillus thuringiensis subsp. aizawai and 0.0009 ppmchlorantraniliprole.

As seen in Table 5, the mixtures of the present invention provided amore than additive effect. By using the following formula, Applicant wasable to determine that this response was synergistic: %Cexp=A+B−(AB/100).

The results of this calculation indicated that the synergy ratio was2.15 at 24 hours and 1.17 at 48 hours. As a finding of higher than 1 isindicative of synergy, the ratios of 1.17 and above are clearlysynergistic. Synergy was shown at a ratio of Bacillus thuringiensissubsp. aizawai to chlorantraniliprole of 1:0.0003.

Example 6 Soybean Looper

In this study, the response of soybean looper larvae to amounts ofBacillus thuringiensis subsp. aizawai and chlorantraniliprole wasobserved. The results of this study are in Table 6.

TABLE 6 Time after % Efficacy treat- Neg. Bta + ment Controlchlorantraniliprole Synergy (h) dH₂O Bta Chlorantraniliprole (Ratio1:0.00003) Ratio 24 0 7 10 22 1.35 48 0 17 13 41 1.48

Bacillus thuringiensis subsp. aizawai was applied at a concentration of27 ppm (27 μg/ml). Chlorantraniliprole was applied at a concentration of0.0009 ppm (0.0009 μg/ml). The Bacillus thuringiensis subsp.aizawai/chlorantraniliprole mixture was applied at a concentration of 27ppm Bacillus thuringiensis subsp. aizawai and 0.0009 ppmchlorantraniliprole.

As seen in Table 6, the mixtures of the present invention provided amore than additive effect. By using the following formula, Applicant wasable to determine that this response was synergistic: %Cexp=A+B−(AB/100).

The results of this calculation indicated that the synergy ratio was1.35 at 24 hours and 1.48 at 48 hours. As a finding of higher than 1 isindicative of synergy, the ratios of 1.35 and above are synergistic.Synergy was shown at a ratio of Bacillus thuringiensis subsp. aizawai tochlorantraniliprole of 1:0.00003.

Example 7 Corn Earworm

In this study, the response of corn earworm larvae to amounts ofBacillus thuringiensis subsp. aizawai and chlorantraniliprole wasobserved. The results of this study are in Table 7.

TABLE 7 Time after % Efficacy treat- Neg. Bta + ment Controlchlorantraniliprole Synergy (h) dH₂O Bta Chlorantraniliprole (Ratio1:0.00003) Ratio 24 0 7 10 17 1.04 48 3 10 7 33 2.02

Bacillus thuringiensis subsp. aizawai was applied at a concentration of27 ppm (27 μg/ml). Chlorantraniliprole was applied at a concentration of0.0009 ppm (0.0009 μg/ml). The Bacillus thuringiensis subsp.aizawai/chlorantraniliprole mixture was applied at a concentration of 27ppm Bacillus thuringiensis subsp. aizawai and 0.0009 ppmchlorantraniliprole.

As seen in Table 7, the mixtures of the present invention provided amore than additive effect. By using the following formula, Applicant wasable to determine that this response was synergistic %Cexp=A+B−(AB/100).

The results of this calculation indicated that the synergy ratio was2.02 at 48 hours. As a finding of higher than 1 is indicative ofsynergy, the finding of a synergy ratio of 2.02 is synergistic. Synergywas shown at a ratio of Bacillus thuringiensis subsp. aizawai tochlorantraniliprole of 1:0.00003.

In summary, synergy was seen against diamondback moth, beet armyworm,soybean looper, corn earworm, cabbage looper and southwestern cornborer.

1. A method of controlling a crop plant pest selected from the groupconsisting of Diamondback moth (Plutella xylostella), Beet armyworm(Spodoptera exigua), Soybean looper (Chrysodeixis includens), Cornearworm (Helicoverpa zea), Cabbage Looper (Trichoplusia ni), andSouthwestern Corn Borer (Diatraea grandiosella) comprising applying asynergistic amount of Bacillus thuringiensis subsp. aizawai andchlorantraniliprole to a plant, wherein the ratio of Bacillusthuringiensis subsp. aizawai to chlorantraniliprole is from about1:0.00003 to about 1:3.
 2. The method of claim 1 wherein the ratio ofBacillus thuringiensis subsp. aizawai to chlorantraniliprole is fromabout 1:0.001 to about 1:1.
 3. The method of claim 2 wherein the ratioof Bacillus thuringiensis subsp. aizawai to chlorantraniliprole is fromabout 1:0.04 to about 1:0.8.
 4. The method of claim 1 wherein the amountof Bacillus thuringiensis subsp. aizawai is from about 50 to about 4,000grams per hectare.
 5. The method of claim 4 wherein the amount ofBacillus thuringiensis subsp. aizawai is from about 100 to about 1,300grams per hectare.
 6. The method of claim 5 wherein the amount ofBacillus thuringiensis subsp. aizawai is from about 150 to about 1,250grams per hectare.
 7. The method of claim 1 wherein the amount ofchlorantraniliprole is from about 20 to about 150 grams per hectare. 8.The method of claim 7 wherein the amount of chlorantraniliprole is fromabout 30 to about 130 grams per hectare.
 9. The method of claim 1wherein crop plant pest is Diamondback moth (Plutella xylostella). 10.The method of claim 1 wherein crop plant pest is Beet armyworm(Spodoptera exigua).
 11. The method of claim 1 wherein crop plant pestis Soybean looper (Chrysodeixis includens).
 12. The method of claim 1wherein crop plant pest is Corn earworm (Helicoverpa zea).
 13. Themethod of claim 1 wherein crop plant pest is Cabbage Looper(Trichoplusia ni).
 14. The method of claim 1 wherein crop plant pest isSouthwestern Corn Borer (Diatraea grandiosella).
 15. The method of claim1 wherein the plant is selected from the group consisting of root andtuber vegetables, bulb vegetables, leafy non-brassica vegetables, leafybrassica vegetables, succulent or dried legumes, fruiting vegetables,cucurbit vegetables, citrus fruits, pome fruits, stone fruits, berry andsmall fruits, tree nuts, cereal grains, forage and fodder grasses andhay, non-grass animal feeds, herbs, spices, flowers, bedding plants,ornamental flowers, artichoke, asparagus, coffee, cotton, tropicalfruits, hops, malanga, peanut, pomegranate, oil seed vegetables,sugarcane, tobacco, turf, and watercress.
 16. The method of claim 15wherein the plant is genetically modified.
 17. The method of claim 15wherein the cereal grains are selected from the group consisting ofbarley, buckwheat, pearl millet, proso millet, oats, corn, field corn,sweet corn, seed corn, popcorn, rice, rye, sorghum (milo), sorghumspecies, grain sorghum, sudangrass (seed), teosinte, triticale, wheat,wild rice, and cultivars, varieties and hybrids thereof.
 18. The methodof claim 17 wherein the plant is genetically modified corn.
 19. Themethod of claim 15 wherein the succulent or dried vegetable legumes areselected from the group consisting of Lupinus beans, Phaseolus beans,Vigna beans, broad beans, chickpea, guar, jackbean, lablab bean, lentil,Pisum peas, pigeon pea, soybean, immature seed soybean, sword bean,peanut, and cultivars, varieties and hybrids thereof.
 20. The method ofclaim 19 wherein the plant is genetically modified soybean.